■Watch on-demand webinar by SS2000 users :
Single Live Cell Mass Spectrometry (SiLC-MS) Analysis
Presenter : Carla Newman (Scientific Leader (Cellular Imaging and Dynamics), GSK)
■Download a whitepaper detailing new research made possible by the SS2000 :
A Novel Method Linking Single Cell Lipidomics to Cellular Function
Professor Melanie Bailey and her group from the University of Surrey have used Yokogawa’s SS2000 to
establish a groundbreaking single-cell LC/MS workflow from live cells.
Subcellular Sampling System
The SS2000 is a system that automatically samples specific regions of cells or whole cells at the single-cell level while imaging cells in culture with a confocal microscope. Because cells in culture do not need to be detached, positional and morphological information is maintained.
Sampling features
- Automated operation.
- Precise pipetting control of location works.
- Sampling with location and morphology information.
- High resolution images and image analysis using confocal microscopy.
- Incubator function for maintaining cell activity.
Use Cases
- Intracellular components can be sampled at a single-cell level. This includes intracellular components that are difficult to sample by biochemical methods, such as organelles without lipid membranes.
- By sampling while maintaining the positional information, it is possible to sample and analyze the normal cells adjacent to the cancer cells and the normal cells located away from the cancer cells.
- Sampling is possible while maintaining morphological information. Allowing cells with different morphological changes can be sampled and compared.
- Different parts of a neuron, such as a cell body or axon, can be targeted for sampling.
- Single-cell cloning is possible from specific cells or cells with specific behavior under microscopic observation, such as transfected cells or virus-resistant cells. By combining various image analysis techniques, accurate and efficient cloning is possible.
- Multiple samples can be collected in the same well for analysis that requires a sample volume.
Details
Subcellular sampling
Unlike existing cell isolation devices, the SS2000 can not only isolate a whole-cell but also sample only the target site inside the cell. It is possible to sample cytoplasm and regions containing target organelles selectively.
After staining of HeLa cell nuclei (blue), cytoplasm (green), and mitochondria (red), a mitochondria-rich region (arrow) of the cytoplasm was sampled.
Maintaining positional and morphological information of cells
Since only the target cells can be sampled without detaching the cells in culture, it is possible to sample while maintaining positional and morphological information.
Normal MDCK cells and green fluorescent-labeled abnormal MDCK cells were co-cultured at a 50:1 ratio. A normal cell adjacent to the abnormal cell exhibiting fluorescent signals (arrows) was sampled.
High usability samples
Samples can be collected on PCR plates and microplates and collect multiple samples in the same well. Samples can also be taken while being held in the glass tip without being ejected. The collection site has a cooling function to suppress sample degradation and an incubator function to maintain the culture environment. These samples can be used for genetic analysis, mass spectrometry, and single-cell cloning.
Live Cell Imaging with Confocal Microscopy
The SS2000 utilizes live-cell imaging products developed by Yokogawa. High-speed, high-resolution 3D imaging is possible using our unique confocal microscope technology. Samples can be taken from targeted cells under a confocal microscope in an incubator environment. Time-lapse photography is also possible, allowing dynamic changes in the target cell to be captured. Since it is possible to record moving images during sampling and images before and after sampling, it is possible to compare the results of analysis of collected samples with cell imaging data.
Target cells and sampling positions can be automatically selected by image analysis. (Targets can be automatically selected as shape of cells, size of nuclei, density of organelles etc.)
Product Specifications
| Automatic sampling functions | Tip diameter | 3μm, 5μm, 8μm, 10μm |
|---|---|---|
| Incubator loader environment | 37℃, 5％CO2, humidified | |
| Collection loader environment | 37℃, 5％CO2, humidified (for culture) / 4℃ (for cooling) | |
| Collection loader compatible vessels | 96-well PCR plate (0.1mL, 0.2mL) Multiwell culture plate (96well) | |
| Postioning precision of sampling | XYZ axial designated resolution: 0.1μm | |
| Imaging functions | Confocal scanning method | Microlens enhanced dual wide Nipkow disk confocal |
| Incubator loader compatible vessels | When sampling cell: φ35mm dishes *1 Microplate (6well, 24well, 96well) |
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| When observing cell: φ35mm dishes *1 Microplate (6well, 12well, 24well, 48well, 96well, 384well, 1536well) Slideglass *2 |
||
| Excitation laser wavelength | 405, 488, 561, 640nm (Uniformizer installed) | |
| Emission filter | Filter size : φ25nm, Maximum slot number : 10 (Electric switching), Adjacent switching speed : 100msec | |
| Transmission illumination | Bright-field, LED source | |
| Objective lens | Dry lens : 4x, 10x, 20x, 40x Long-working distance lens:20x, 40x Note that only the 40x dry lens can be used for cell sampling. |
|
| Z focus | Electric Z motor, designated resolution : 0.1μm | |
| Electric stage | XYZ axial designated resolution : 0.1μm | |
| Autofocus | Laser autofocus | |
| Camera | sCMOS camera 2,000 x 2,000pixel Pixel size : 6.5 x 6.5μm | |
| Other | Special purpose workstation | Workstation for sampling, measurement, analysis, 24 inch display x2 |
| Measurement software | Measurement functions (2D, 3D, Time-lapse, Map imaging), Viewing measurement and sampling data, Reporting functions (Image data, Video data), Whole cell sampling, Intracellular component sampling | |
| Analysis software | Analysis functions (3D, Tile, Label-free, Texture analysis, Deep Learning, Gating), 3D viewer, Graphing functions, Reporting functions (Image data, Video data, EC50, IC50, Z'-factor) | |
| External dimensions, Weight | Main unit : W1,217 x D643 x H595 mm, 145kg Utility box : W275 x D432 x H298 mm, 18kg Gas mixer : W275 x D432 x H298 mm, 10kg Special purpose workstation : W172 x D471 x H414 mm, 14kg Display : W531 x D500 x H166 mm, 5.6kg |
|
| Operating environment | Temperature : 15 to 30℃ Humidity : 30 to 70％RH no condensation |
|
| Power consumption | Main unit, Utility box and Gas mixer : 1,200VAmax Workstation : 950VAmax Display : 42VAmax x 2 |
|
| Data formats (Measurement software) | Captured images : 16bit TIFF (OME-TIFF, TIFF) Output image data : TIFF, PNG, JPEG Output video data : WMV, MPEG4 |
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Data formats (Analysis software) |
Numeric data : CSV Output image data : TIFF, PNG, JPEG Output video data : WMV, MPEG4 |
*1 A sample holder is required, and with it, up to 3 samples can be installed.
*2 A sample holder is required, and with it, up to 4 samples can be installed.
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Resources
In recent years, research on single cells has become increasingly popular, but with more sensitive analytical techniques, it has become possible to analyze specific intracellular components such as organelles at the single-cell level.
The SS2000 is an innovative system that can sample intracellular components at the single-cell level using a tip (glass capillary) with a diameter of several micrometers while imaging with a confocal microscope. In this application note, we introduce a case in which intracellular components were sampled by SS2000 and genetic analysis was performed by qPCR.
In recent years, single-cell studies have become increasingly popular as analytical techniques have become more sensitive and widespread.
The SS2000 is a revolutionary device that can sample intracellular components and single cells using glass capillary tips with a diameter of several micrometers while performing confocal microscope imaging. These application notes introduce examples of sampling for various cell types.
In recent years, single cell analysis has become increasingly popular due to the new development of advanced and high sensitivity analytical methods.
SS2000 is a revolutionary system that can sample subcellular components and a whole cell by using a glass capillary tip with an inner diameter of a few μm while imaging with a confocal microscope.
This application note provides an example of single cell RNA sequencing scRNA seq) from cells sampled by SS2000, and the data is comparable to conventional methods.
In recent years, research on single cells has become increasingly popular. With more sensitive analytical techniques, it has become possible to analyze specific intracellular components such as organelles, even at the single-cell level.
The SS2000 is an innovative system that can sample subcellular intracellular components using a glass capillary having a tip diameter of only a few micrometers while imaging with a confocal microscope.
In this application note, we introduce a case where after drug treatment, intracellular components were sampled by the SS2000 and analyzed by single-cell mass spectrometry.
The SS2000 is a revolutionary system that allows both confocal imaging and sampling of targeted intracellular components or single cells using a glass tip with an inner diameter of a few micrometers. The SS2000 includes an incubator as well as highcontent imaging functions, such as long time-lapse observation, machine learning, and label-free analysis.
The SU10 is an innovative device that delivers target substances directly into cells or into nuclei with a nanopipette that has a tip outer diameter as small as a few tens nm.
This application note shows a case study of sampling and analysis with the SS2000 from cells delivered intracellularly with the SU10.
In recent years, single-cell studies have become increasingly popular, leading to the development of instruments capable of spatial omics - the integration of samples' 3D locations and their omics data. The SS2000 is a revolutionary device that can collect intracellular components at the single-cell level and image the samples using a confocal microscope. In this application note we introduce the research performed by Dr. Okada of The University of Tokyo, who established an intra single living cell sequencing (iSC-seq) method which uses SS2000 to sample intracellular components from osteoclasts (a type of multinucleated giant cell) and analyzes them through next-generation sequencing.
In recent years, research on single cells has been actively conducted.However, with more sensitive analytical techniques, it is now possible to analyze specific intracellular components at the single-cell level.
Videos
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News
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News Brief Apr 26, 2024 Yokogawa Helps to Revolutionize the Field of Single-Cell Lipidomics
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Press Release Dec 1, 2021 Yokogawa Develops Single Cellome System SS2000 for Subcellular Sampling
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